Functional role of PLOD3 and LNP delivery of siPLOD3 in pancreatic cancer

Background: Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase 3 (PLOD3) is a gene that encodes a protein crucial for the formation of hydroxylysine in collagens. Overexpression of PLOD3 has been identified in various types of cancer, suggesting its potential role in promoting tumor growth and metastasis. Lipid nanoparticles (LNPs) represent a rapidly evolving method for delivering therapeutic agents and have been the focus of extensive research for enhancing targeted delivery of therapeutic agents to tumors. The goal of my thesis study is to investigate the functional role of PLOD3 and the use of LNPs for delivery of PLOD3 siRNA (siPLOD3-LNPs) in pancreatic cancer.Methods: Analysis of PLOD3 gene expression in pancreatic adenocarcinoma (PAAD) was conducted using the data from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) databases. In addition to these databases, western blot and immunohistochemistry (IHC) were used to assess the expression of the PLOD3 gene in PAAD. In order to discover the role of PLOD3 and evaluate the use of siPLOD3-LNP in pancreatic cancer, CCK-8, migration, Matrigel invasion, and colony-forming assays were carried out on MIA PaCa-2, BxPC-3, and KPC pancreatic cancer cell lines. The synergistic effect of cisplatin and siPLOD3-LNP on pancreatic cancer cell proliferation was also assessed with the CCK-8 assay. Additionally, RNA- sequencing was performed to investigate the changes in global gene expression following siPLOD3-LNP treatment. Results: Based on statistical analysis of the data from the GEO and TCGA databases and the result from IHC, it was observed that PLOD3 gene expression was significantly increased in PAAD tumor tissues compared to normal controls. Moreover, western blot analysis also showed a higher expression level of PLOD3 in pancreatic cancer cells compared to normal pancreatic cells. Western blot analysis demonstrated a significant reduction in PLOD3 protein expression in the pancreatic cancer cells treate